Retinal pigment epithelium (RPE) cells are vital for retinal health. However, they are susceptible to injury with ageing and exposure to excessive light, including UV (100-380 nm) and visible (380-760 nm) radiation. To evaluate the protective effect of blueberry anthocyanins on RPE cells, in vitro cell models of replicative senescent and light-induced damage were established in the present study. After purification and fractionation, blueberry anthocyanin extracts (BAE) were yielded with total anthocyanin contents of 31·0 (SD 0·5) % and were used in this study. Replicative senescence of RPE cells was induced by repeatedly passaging cells from the fourth passage to the tenth. From the fifth passage, cultured RPE cells began to enter a replicative senescence, exhibiting reduced cell proliferation along with an increase in the number of β-galactosidase-positive cells. RPE cells maintained high cell viability (P < 0·01) and a low (P < 0·01) percentage of β-galactosidase-positive cells when treated with 0·1 μg/ml BAE. In contrast, after exposure to 2500 (SD 500) lx light (420-800 nm) for 12 h, RPE cells in the positive control (light exposure, no BAE treatment) exhibited premature senescence, low (P < 0·01) cell viability and increased (P < 0·01) vascular endothelial growth factor (VEGF) release compared with negative control cells, which were not subjected to light irradiation and BAE exposure. Correspondingly, BAE is beneficial to RPE cells by protecting these cells against light-induced damage through the suppression of ageing and apoptosis as well as the down-regulation of the over-expressed VEGF to normal level. These results demonstrate that BAE is efficacious against senescence and light-induced damage of RPE cells.
Studies suggest that the consumption of berry fruits rich in anthocyanins may have beneficial effects on improving visual function. This study determined the total polyphenol and total anthocyanin contents in wild Chinese blueberries using the Folin-Ciocalteu reagent method and a pH differential method. Anthocyanin composition and quantity were characterized by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry coupled with mass selective detection. Total polyphenol and anthocyanin contents were 602.9 ± 9.2 and 177.8 ± 8.3 mg/100 g, respectively. Seventeen anthocyanins were discovered, and only 13 were tentatively identified in the wild blueberries. Anthocyanins of malvidin glycosylated with hexose or pentose accounted for >46% of total anthocyanin content. Furthermore, the effect of whole blueberries on retinal damage in pigmented rabbits upon light exposure was investigated, and the retinal functions were evaluated by electroretinogram (ERG). Pigmented rabbits were chosen for this experiment because of their large eyes, which facilitated the operative procedure and observation, and the similarity of their eyes to the human eye structure. Light-induced retinal damage was induced by exposure to white light at 15000 ± 1000 lx for 2 h. Feeding the rabbits with blueberries at a dosage of 1.2 or 4.9 g/kg/day for 4 weeks prior to light exposure effectively reduced photodamage to the retinas. This study adds to the growing body of data supporting the bioactivity of blueberries in improving mammal vision.
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